title: Towards a recombinant vaccine against diphtheria toxin.
abstract: Two recombinant fragments of diphtheria toxin (DT) were fused to an engineered tandem repeat of the immunoglobulin (Ig) binding domain of protein A, called ZZ. These fragments are (i) the receptor binding domain (DTR), which comprises amino acids 382 to 535 of DT, and (ii) a linear peptide (DT(168-220)) which comprises residues 168 to 220 of the loop between fragment A and fragment B of DT. The fusion proteins were produced in Escherichia coli and purified by affinity chromatography. In vitro experiments showed that the DTR domain is responsible for the capacity of ZZ-DTR to bind to Vero cells and is capable of inhibiting the cytotoxicity of DT for these cells. These findings suggest that DTR binds to the cell surface receptors of DT and hence adopts a conformation that is similar to that of the receptor binding domain of DT. We compared the capacities of ZZ-DTR, ZZ-DT(168-220), and a chemically detoxified form of DT currently used for vaccination to elicit antibodies in rabbits. The toxoid was more immunogenic than ZZ-DT(168-220), which in turn was more immunogenic than ZZ-DTR. However, ZZ-DT(168-220) antiserum was poorly efficient at neutralizing DT cytotoxicity on Vero cells, whereas ZZ-DTR antiserum was only 15-fold less potent than anti-DT antisera.
authors.affiliation: CEA, Départment d'Ingénierie et d'Etudes des Protéines, Centre d'Etudes de Saclay, Gif-sur-Yvette, France.
authors.forename: K
authors.initials: K
authors.lastname: Lobeck
authors.forename: P
authors.initials: P
authors.lastname: Drevet
authors.forename: M
authors.initials: M
authors.lastname: Léonetti
authors.forename: C
authors.initials: C
authors.lastname: Fromen-Romano
authors.forename: F
authors.initials: F
authors.lastname: Ducancel
authors.forename: E
authors.initials: E
authors.lastname: Lajeunesse
authors.forename: C
authors.initials: C
authors.lastname: Lemaire
authors.forename: A
authors.initials: A
authors.lastname: Ménez
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D004167:Diphtheria Toxin / Q000276:immunology*; D004168:Diphtheria Toxoid / Q000276:immunology*; D008297:Male; D009500:Neutralization Tests; D010446:Peptide Fragments / Q000276:immunology*; D011817:Rabbits; D014614:Vaccines, Synthetic / Q000276:immunology*; D014709:Vero Cells
chemical_list: D004167:Diphtheria Toxin; D004168:Diphtheria Toxoid; D010446:Peptide Fragments; D014614:Vaccines, Synthetic